Validated high-performance thin-layer chromatographic analysis of ursolic acid and β-sitosterol in the methanolic fraction of<i>Paederia foetida</i>L. leaves

Abstract

A high-performance thin-layer chromatography (HPTLC) method for the simultaneous quantitative determination of ursolic acid and β-sitosterol in the methanolic fraction of Paederia foetida L. leaves was developed for the first time. For achieving good separation, a mobile phase of toluene‒ethyl acetate‒formic acid (8:2:0.1, v/v) was used. The densitometric determination was carried out at 550 and 522 nm in reflection/absorption mode for ursolic acid and β-sitosterol. The calibration curves were linear in the range of 100-600 ng per spot for ursolic acid and β-sitosterol. During the analysis, the methanolic fraction of P. foetida L. leaves showed the presence of ursolic acid (0.12 ± 0.05%) and β-sitosterol (0.08 ± 0.12%). The proposed method is simple, precise, specific, accurate, less time-consuming, and cost-effective. The statistical analysis of the data obtained proves that the method is reproducible and selective and can be used for the routine analysis of the reported phenolic compounds in crude drug and extracts. The simultaneous quantification of these compounds has not yet been reported in P. foetida L. leaves which may be utilized for the proper standardization of the plant.